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NuMA-related LIN-5, ASPM-1, calmodulin and dynein promote meiotic spindle rotation independently of cortical LIN-5/GPR/Gα

Nature Cell Biology volume 11, pages 269–277 (2009)Cite this article

Abstract

The spindle apparatus dictates the plane of cell cleavage, which is critical in the choice between symmetric or asymmetric division. Spindle positioning is controlled by an evolutionarily conserved pathway, which involves LIN-5/GPR-1/2/Gα in Caenorhabditis elegans, Mud/Pins/Gα in Drosophila and NuMA/LGN/Gα in humans1. GPR-1/2 and Gα localize LIN-5 to the cell cortex, which engages dynein and controls the cleavage plane during early mitotic divisions in C. elegans2,3,4,5,6. Here we identify ASPM-1 (abnormal spindle-like, microcephaly-associated) as a novel LIN-5 binding partner. ASPM-1, together with calmodulin (CMD-1), promotes meiotic spindle organization and the accumulation of LIN-5 at meiotic and mitotic spindle poles. Spindle rotation during maternal meiosis is independent of GPR-1/2 and Gα, yet requires LIN-5, ASPM-1, CMD-1 and dynein. Our data support the existence of two distinct LIN-5 complexes that determine localized dynein function: LIN-5/GPR-1/2/Gα at the cortex, and LIN-5/ASPM-1/CMD-1 at spindle poles. These functional interactions may be conserved in mammals, with implications for primary microcephaly.

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Figure 1: LIN-5 and CMD-1 associate with ASPM-1.
Figure 2: Localizations of LIN-5 and DHC-1 dynein heavy chain to meiotic spindle poles are dependent on ASPM-1 and CMD-1.
Figure 3: Localization of LIN-5 at the spindle poles, but not at the cortex, is dependent on ASPM-1 and CMD-1.
Figure 4: ASPM-1 is not required for cortical pulling forces and spindle positioning in early mitotic divisions.
Figure 5: LIN-5, ASPM-1, CMD-1 and DHC-1 are required for spindle rotation during meiosis.

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Acknowledgements

We are grateful to Steve Gygi and Ross Tomaino (Taplin Biological Mass Spectrometry, Harvard Medical School) for analysis of peptides by mass spectrometry (HMS), to Sean O'Rourke and Bruce Bowerman for the GFP–DHC-1 strain EU1561, to Inge The for assistance with ASPM-1 antibody production, Kristiaan de Vries for scoring embryonic lethality, and the Caenorhabditis Genetics Center (National Institutes of Health, National Center for Research Resources) for providing strains. We thank Adri Thomas and Marjolein Wildwater for critically reading the manuscript. This work was supported in part by National Institutes of Health grant GM57990 to S.v.d.H, Swiss National Science Foundation grant 3100A0-102087 to P.G., and Marie Curie International Reintegration grant MIRG-CT-2007-046458 to M.B.

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  1. Audrey Perreault

    Present address: Present address: Department of Biochemistry, Université de Sherbrooke, Québec, Canada.,

Authors and Affiliations

  1. Developmental Biology, Utrecht University, Padualaan 8, Utrecht, 3584, CH, The Netherlands

    Monique van der Voet, Christian W. H. Berends, Mike Boxem & Sander van den Heuvel

  2. Swiss Institute for Experimental Cancer Research (ISREC), Swiss Federal Institute of Technology (EPFL), School of Life Sciences, Lausanne, CH-1066, Switzerland

    Tu Nguyen-Ngoc & Pierre Gönczy

  3. Center for Cancer Systems Biology (CCSB) and Department of Cancer Biology, and Department of Genetics, Dana-Farber Cancer Institute, Harvard Medical School, Boston, 02115, Massachusetts, USA

    Marc Vidal & Mike Boxem

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Contributions

M.v.d.V. was primarily responsible for the experiments, data analysis and manuscript writing. C.B. performed IP-western experiments and fusion protein purification. A.P. contributed in the initial phase of the project. T.N-N. and P.G. performed spindle severing assays. M.B. performed Y2H experiments with support from M.V. S.v.d.H. contributed to experimental design, supervision and manuscript writing.

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Correspondence to Sander van den Heuvel.

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van der Voet, M., Berends, C., Perreault, A. et al. NuMA-related LIN-5, ASPM-1, calmodulin and dynein promote meiotic spindle rotation independently of cortical LIN-5/GPR/Gα. Nat Cell Biol 11, 269–277 (2009). https://doi.org/10.1038/ncb1834

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